Search

São Paulo state is one of the country’s largest producers of beef and milk, and the midwestern region plays a key role in this production, as half of São Paulo’s cattle herd is found in this region. These numbers alone demonstrate the importance of livestock in this region. Therefore, this study aimed to describe the main epidemiological and clinical signs in cattle cases at the Large Animal Hospital at FMVZ-Unesp, located in the midwestern region of São Paulo state. The present retrospective study assessed 638 clinical cases of cattle treated from January 2010 to December 2019 (10 years). Digestive system diseases were the most prevalent, diagnosed in 30.3% of patients, followed by neurological diseases (19.1%) and respiratory diseases (10.5%). The other diseases were distributed in decreasing order as follows: musculoskeletal (7.8%), hematopoietic (6.1%), genitourinary (5.6%), metabolic and nutritional (5.5%), neonatal (4.7%), cutaneous (2.6%), poisoning (2.5%), lymphatic (2.2%), cardiovascular (1.6%) and other diseases (1.4%). Rabies, a fatal zoonotic disease, was the main cause of death in this study and the main disease associated with neurological signs (23.7%). Recognizing the main diseases of cattle in this region will promote the adoption of prophylactic measures to minimize their occurrence and manage treatment to avoid economic losses and decreased productivity of herds. RESUMO: O estado de São Paulo é um dos maiores produtores de carne e leite bovino do país e a região centro-oeste do estado tem papel fundamental nessa produção, pois possui a metade do rebanho de bovinos do estado. Portanto, este estudo descreve os principais achados clínicos e epidemiológicos de bovinos atendidos no Hospital de Grandes Animais da FMVZ/Unesp, localizado na região centro-oeste paulista. Foi realizado um levantamento nos arquivos da Clínica de Grandes Animais do Hospital Veterinário da FMVZ-Unesp, Botucatu/SP, dos 638 casos clínicos de bovinos atendidos de janeiro de 2010 a dezembro de 2019. Enfermidades do sistema digestório foram as mais prevalentes (30,3%), seguidas das doenças neurológicas (19,1%) e respiratórias (10,5%). As demais enfermidades foram distribuídas, em ordem decrescente, em: musculoesquelético (7,8%), haematopoiético (6,1%), geniturinário (5,6%), cutânea (2,6%), linfático (2,2%) e cardiovascular (1,6%) ou foram classificadas em doenças metabólicas e nutricionais (5,5%), neonatais (4,7%), tóxicas (2,5%) ou outros distúrbios (1,4%). A raiva, uma zoonose fatal, foi a principal causa de óbito neste estudo e a principal doença neurológica (23,7%). O reconhecimento das principais doenças dos bovinos desta região permite a adoção de medidas profiláticas e de manejo para minimizar sua ocorrência e evitar perdas econômicas com o tratamento e menor produtividade do rebanho.

Made available in DSpace on 2022-04-29T08:35:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Background: In the Quarter Horse (QH), myosin heavy chain myopathy (MYHM), which is characterised by nonexertional rhabdomyolysis or immune-mediated myositis (IMM) with acute muscle atrophy, is strongly associated with the missense E321G MYH1 mutation. Objectives: To document the existence of MYHM in the Brazilian QH population, this study includes a case report of two related QH foals with the E321G MYH1 mutation that had clinical signs of MYHM, with histological confirmation of IMM in one of the foals. This prompted an investigation the aim of which was to determine the allele frequency of the E321G MYH1 variant across QHs using a DNA archive in Brazil. Study design: Cross sectional. Methods: To estimate the allele frequency of the E321G MYH1 variant in Brazilian QHs, 299 DNA samples from QHs used in different disciplines (reining, barrel racing, halter, cutting and racing) were analysed. DNA fragments containing the region with the mutation were amplified by PCR and used for direct genomic sequencing. Results: Of the 299 genotyped QHs, 44 animals (14.7%) were heterozygous (My/N) for the E321G MYH1 variant, and 255 (85.3%) were homozygous for the wild-type allele (N/N), implying an allele frequency of 0.074. Reining horses had a significantly higher prevalence of heterozygosity than horses in other disciplines (P =.008). Main limitations: The DNA samples were collected from 2010 to 2014. As only registered QHs were evaluated, the results may not reflect the actual incidence in the general population of Brazilian QHs. Conclusions: The reported cases of MYHM and the high prevalence of the MYH1 mutation found in the assessed Brazilian QH population, particularly in reining QHs, suggests that MYHM should be included in genetic screening. Reasonable control measures are important to prevent an increase in the incidence of MYHM in QHs in Brazil. Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) School of Veterinary Medicine Universidade Federal de Uberlândia Sector of Veterinary Pathology Veterinary Faculty Federal University of Rio Grande Sul Department of Large Animal Clinical Sciences College of Veterinary Medicine Michigan State University Department of Population Health and Reproduction School of Veterinary Medicine University of California-Davis Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)

Hereditary myotonia (HM) is characterized by delayed muscle relaxation after contraction as a result of a mutation in the CLCN1 gene. We describe here a complex CLCN1 variant in a mixed-breed dog with clinical and electromyographic signs of HM. Blood samples from the myotonic dog, as well as from his male littermate and parents, were analyzed via amplification of the 23 exons encoding CLCN1. After sequencing the CLCN1 gene, a complex variant was found in exon 6 c.[705T>G; 708del; 712_732del], resulting in a premature stop codon in exon 7 and a protein that was 717 amino acids shorter than the normal CLC protein. The myotonic dog was identified as homozygous recessive for the complex CLCN1 variant; its parents were heterozygous, and its male littermate was homozygous wild-type. Knowledge of the CLCN1 mutations responsible for the development of hereditary myotonia allows greater clarification of this condition.

Made available in DSpace on 2022-05-01T10:19:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Analysis of the cerebrospinal fluid (CSF) is important as a complementary test in horses with neurologic diseases, and sequential analysis may provide information about the treatment response or evolution and quantitative measures of the CSF drug concentration during treatment. The aim of this study was to compare erythrocyte and nucleated cell counts and protein concentration in multiple CSF samples obtained sequentially from two different puncture sites in clinically healthy horses. Eight and 12 horses, with no evidence of neurologic disease, were subjected to CSF collection from the atlanto-occipital (AO) and C1–C2 spaces, respectively. Cytologic and chemical analyses were performed on the CSF obtained at five sampling times (T1, T2, T3, T4, and T5). Repeated measures models were used to compare the mean erythrocyte count, nucleated cell count, and total protein concentration between the AO and C1–C2 groups at each sampling time. C1–C2 CSF had a significantly higher total protein concentration at T1 and T4 than that of AO CSF. All total protein concentration values remained within the reference interval (

Equine leukoencephalomalacia (LEM) is a disease caused by the ingestion of food, especially corn, contaminated by fumonisin, a Fusarium verticillioides (synonymous with F. moniliforme) metabolite. The clinical signs of brain injuries have an acute onset and rapid evolution. This study aimed to describe the clinical findings in 11 animals diagnosed with LEM, including cerebrospinal fluid (CSF) analysis. Of these animals, 91% (10/11) were horses, and only 9% (1/11) were asinine. The clinical localization of the lesions was 64% (7/10) cerebral, manifested mainly by altered mental state and behavioral disturbance, and 36% (4/11) were brainstem lesions, manifested by incoordination, head tilt, nystagmus, facial hypoalgesia, difficulty in apprehension, chewing, and swallowing food. Postmortem findings revealed that 82% (9/11) of the lesions were in the cerebrum and 18% (2/11) in the brainstem. CSF findings, such as xanthochromia (43%, 3/7), hyperproteinorrachia (50%, 3/6), and pleocytosis (43%, 3/7) were observed. The affected animals showed neurological signs that were compatible with cerebral and/or brainstem injuries. The CSF from animals with LEM may present with xanthochromia, hyperproteinorrachia, and pleocytosis, reinforcing the fact that this disease should be included in the differential diagnosis of encephalomyelopathies. RESUMO: A leucoencefalomalácia (LEM) é uma enfermidade que acomete equídeos causada pela ingestão de milho e seus derivados e feno contaminados pela micotoxina fumonisina, um metabólito do fungo Fusarium verticillioides (sinônimo para F. moniliforme). Os sinais clínicos apresentam início agudo e evolução rápida e são decorrentes de lesões encefálicas. O objetivo deste estudo é descrever os achados clínicos de 11 equídeos diagnosticados com LEM, incluindo a análise do líquido cefalorraquidiano (LCR). 91% dos animais afetados eram equinos e somente 9% (1/11) era asinino. A localização clínica das lesões era 64% (7/10) cerebrais, manifestadas por alterações no estado mental e comportamento e 36% (4/10) no tronco encefálico, manifestadas por incoordenação, desvio lateral de cabeça, nistagmo, hipoalgesia da face e dificuldade de apreensão, mastigação e deglutição de alimentos. Comparativamente, os achados post mortem revelaram que 82% (9/11) das lesões eram no cérebro e 18% (2/11) no tronco encefálico. Alterações no LCR, tais como xantocromia (43%, 3/7), hiperproteinorraquia (50%, 3/6) e pleocitose (43%, 3/7), foram observadas. Os animais afetados apresentaram sinais clínicos compatíveis com lesões encefálicas e/ou de tronco cerebral. O LCR de animais com LEM pode apresentar xantocromia, hiperproteinorraquia, e pleocitose, reforçando que esta doença deve ser incluída como diagnóstico diferencial de encefalomielites.

The Labrador Retriever is among the main breeds with the greatest predisposition to obesity. Several factors, especially the interrelationships between food management, exercise and social factors; influence the likelihood of a dog becoming obese. Furthermore, genetic factors are also responsible for obesity in dogs, and in Labrador Retriever, a frameshift mutation (P187fs) in pro-opiomelanocortin (POMC) gene is strongly associated with obesity. There is no knowledge of studies that have previously evaluated the prevalence of the canine POMC deletion (P187fs) in Brazilian Labrador Retriever. Therefore, the objective of this study was to investigate this mutation in Labrador Retriever dogs in Brazil. Of the 108 Labrador Retrievers that were assessed in this study, 59 were from a previous study, composed by animals assisted in a veterinary hospital with unknown lineage, and 49 were from a prospective study, composed of 19 pet and 30 assistance/rescue Labrador Retriever dogs. The obesity risk and appetite questionnaire were applied, with some modifications, to tutors of the animals used in the prospective study. Fragments of the DNA, containing the mutation, were amplified by PCR and submitted to direct gene sequencing. The allele frequency of the mutation was 21.3% and was out of Hardy-Weinberg equilibrium (P0.05). Although the mutation has increased the food-motivation in the assistance/rescue dogs, other variables, especially frequent exercising, favored that these animals maintained the ideal body weight (body condition score = 5). In summary, the Hardy-Weinberg disequilibrium observed in the allele distribution of the deletion POMC_P187fs in this study, independently of the Labrador Retriever group assessed, suggesting the possibility of positive selection of the mutated allele, which may lead to the maintenance of this deleterious allele in the studied population. RESUMO: O Labrador Retriever é uma das principais raças caninas com maior predisposição à obesidade. Vários fatores, especialmente as interrelações entre a alimentação, exercício e fatores sociais, influenciam a probabilidade de um cão se tornar obeso. Além disso, fatores genéticos são também responsáveis pela obesidade em cães, e no Labrador Retriever a mutação “frameshift” P187fs no gene pró-opiomelanocortina (POMC) está fortemente associada à obesidade. Não existem estudos prévios de prevalência da deleção P187fs no gene POMC em cães Labrador Retriever no Brasil. Portanto, o objetivo deste estudo foi investigar esta mutação em cães da raça Labrador Retriever no Brasil. Dos 108 Labradores Retrievers avaliados neste estudo, 59 eram de um estudo retrospectivo (composto por animais atendido no hospital veterinário e sem linhagem conhecida) e 49 eram de um estudo prospectivo (composto por 19 cães pet e 30 cães de assistência/resgate). Um questionário de risco de obesidade modificado foi aplicado nos tutores dos animais usados no estudo prospectivo. Fragmentos de DNA, contendo a mutação, foram amplificados por PCR e submetidos ao sequenciamento gênico direto. A frequência alélica da mutação foi de 21,3% e estava fora do equilíbrio de Hardy-Weinberg (P

Made available in DSpace on 2021-06-25T11:01:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 Dwarfism is a skeletal disorder that causes abnormal growth. In Miniature horses, dwarfism can occur as chondrodysplastic dwarfism, an autosomal recessive disorder associated with five mutations (D1, D2, D3*, D4 and c.6465A > T variant) in the aggrecan (ACAN) gene. The aim of this study was to evaluate the expression of aggrecan (at the gene and protein level) and specific cytokines (IL-1β, IL-6, and TNF-α) in the articular cartilage of Miniature horses with chondrodysplastic dwarfism (D4/c.6465A > T genotype). Metatarsal bone samples from eight dwarf Miniature horses were collected for histopathological analysis, and articular cartilage was collected to detect and quantify aggrecan levels through Western blotting and determine the relative expression levels of ACAN, IL-1β, IL-6, and TNF-α through qPCR. All affected animals presented chondrodysplasia-like lesions with disorganization of the chondrocyte layers and reduced the amount of an extracellular matrix. No significant difference in aggrecan expression levels in uncleaved samples from the dwarf and control groups (composed of phenotypically normal animals of similar age and breed (P =.7143)) was found using Western blotting. qPCR revealed that ACAN gene expression was higher in the affected animals than in normal animals (P =.0119). No significant difference in cytokine levels was detected between the groups. Mutant aggrecan may interfere with normal cellular function, leading to chondrodysplasia and the observed phenotypic findings. São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science

Made available in DSpace on 2022-04-29T08:36:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 The visual evoked potential (VEP) has many applications in veterinary neurology, but the test is not routinely used in a clinical setting. The aim of this study was to describe a reliable method for recording flash visual evoked potentials (F-VEPs) in nonsedated horses. F-VEPs were recorded from both eyes in 20 healthy and calm, adult horses. Recordings were accomplished without sedation, anaesthesia, or the use of mydriatic drugs. The mean and standard deviation of the latency of the most evident positive peak was 52.76±2.37 ms (P53). The mean latencies of the preceding and following negative peaks were 38.14±4.62 (N38) and 72.35±5.33 ms (N72), respectively. There were 2 mean peak-to-peak amplitudes (N38 – P53 and P53 – N72), and they were 11.85±6.21 and 22.81±11.50 µV, respectively. F-VEP was also recorded from 3 horses (6 eyes) before and during sedation with 2 doses each of xylazine (0.4 and 1.1 mg/kg) or detomidine (0.005 and 0.014 mg/kg). It was possible to obtain a reliable F-VEP with a P53 latency in horses without sedation that was similar to the P2 peak described in previous studies, and these data can be used in the future as a normal reference for comparisons in horses with different diseases using a similar methodology. Sedation affected the results by depressing peak amplitudes and increasing latencies or by completely obscuring any response. The exact impact of sedation on VEPs must be evaluated with much caution due to the small sample size. Faculdade de Medicina Veterinária e Zootecnia Universidade Federal de Mato Grosso do Sul Department of Neurology Psychology and Psychiatry College of Medicine - Univ Estadual Paulista (UNESP) Botucatu Department of Veterinary Clinical Science College of Veterinary Medicine and Animal Science - Univ Estadual Paulista (UNESP), Botucatu Department of Neurology Psychology and Psychiatry College of Medicine - Univ Estadual Paulista (UNESP) Botucatu Department of Veterinary Clinical Science College of Veterinary Medicine and Animal Science - Univ Estadual Paulista (UNESP), Botucatu

Made available in DSpace on 2021-06-25T10:56:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-12 Congenital erythropoietic porphyria (CEP) is a rare hereditary autosomal recessive disease which has never been reported in Zebu cattle. A 3-day-old Gir calf showed teeth discoloration, fever, dehydration, and dyspnea. The main gross findings were pink-colored teeth, red-brown periosteum and bone marrow, and a fluorescent bright pink coloration of the bone marrow and articular surfaces under ultraviolet light. Aggregates of periodic acid-Schiff (PAS)-stained porphyrin pigments were evident in the lungs, kidneys, and the liver. An intron 8 single-nucleotide polymorphism (SNP) in both the Gir calf and control animals, along with the absence of the uroporphyrin III synthetase (UROS) gene mutation, was observed. Most SNPs were located in the intron regions of the UROS gene without relevance for CEP. A continuous loss of genetic variability and an increase in inbreeding in some herds may be related to CEP in Gir cattle, one of the most prominent Zebu breeds worldwide. In summary, this study describes a presumptive case of CEP in a Gir calf based on clinical and pathological findings. A definitive diagnosis would require the measurement of porphyrin levels in blood, urine, or tissues or the identification of UROS gene defects. Veterinary Pathology Laboratory University of Brasília, Campus Darcy Ribeiro Instituto Nacional de Investigación Agropecuaria Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Department of Veterinary Clinics and Surgery Veterinary College Universidade Federal de Minas Gerais College of Agronomy and Veterinary Medicine Large Animal Veterinary Teaching Hospital University of Brasília Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)

Made available in DSpace on 2021-06-25T10:15:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Background: Paraoxonase-1 (PON-1) is an antioxidant enzyme, whose activity decreases during the acute phase response in many species. Little is known about PON-1 and its role as a negative acute phase protein during septic inflammation in horses, but promising findings about its utility in diagnosing SIRS and predicting the outcome in diseased horses, were recently highlighted. The objective of the study was to investigate the behaviour of PON-1 in horses after experimentally induced endotoxemia. To this aim, PON-1 activity was measured on 66 plasma samples collected from six clinically healthy mares, previously included in another study, before and at multiple time points between 12 and 240 h after intravenous infusion of Escherichia coli O55:B5 lipopolysaccharide (LPS). Results: Compared with baseline values, a progressive transient decrease of PON-1 activity was observed starting from 24 h post-infusion, with lowest values observed between 3 to 7 days post-infusion, followed by a normalisation to pre-infusion levels the tenth day. Conclusions: The results of this study suggest that measurement and monitoring of PON-1 activity might be useful to evaluate progression and recovery from endotoxemia in horses. Further studies in horses with naturally occurring sepsis are warranted. Department of Veterinary Medicine Veterinary Teaching Hospital University of Milan Department of Veterinary Sciences Veterinary Teaching Hospital Univeristy of Pisa Department of Veterinary Clinical Science School of Veterynary Medicine and Animal Sicence Sao PAulo State University (Unesp) School of Veterinary Science Veterinary Teaching Hospital University of Sydney Department of Veterinary Clinical Science School of Veterynary Medicine and Animal Sicence Sao PAulo State University (Unesp)

Made available in DSpace on 2020-12-12T02:49:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Chondrodysplastic dwarfism in Miniature horses is an autosomal recessive disorder previously associated with four mutations (D1, D2, D3*, and D4) in the aggrecan (ACAN) gene. The aim of this study was to identify additional variants in the candidate ACAN gene associated with chondrodysplastic dwarfism in Miniature horses. Fifteen dwarf Miniature horses were found to possess only one of the dwarfism-causing variants, and two possessed none of the variants. The ACAN exons (EquCab3.0) of seven dwarf Miniature horses were sequenced. A missense SNP in coding exon 11 (g.95271115A > T, c.6465A > T—RefSeq XM_005602799.2), which resulted in the amino acid substitution p.Leu2155Phe (RefSeq XP_005602856.2), was initially associated with the dwarf phenotype. The variant was tested and found present in 14 dwarf foals as well as one parent of each, and both parents of a dwarf possessing two copies. Genetic testing of 347 phenotypically normal Miniature horses demonstrated that none had more than one of the dwarf alleles or c.6465A > T. However, a study of large breeds revealed the presence of c.6465A > T, which was present in homozygosis in two Mangalarga Marchador horses. We suggest that c.6465A > T as a marker of disequilibrium or complex interactions in the Miniature horse genome could contribute to the associated dwarfism. School of Veterinary Medicine and Animal Science São Paulo State University (Unesp) Institute for Biotechnology São Paulo State University (Unesp) School of Agriculture São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science São Paulo State University (Unesp) Institute for Biotechnology São Paulo State University (Unesp) School of Agriculture São Paulo State University (Unesp) FAPESP: 2016/24025-0 FAPESP: 2016/24767-7 FAPESP: 2018/11365-3 CNPq: 307686/2018-3

Made available in DSpace on 2021-06-25T12:28:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Hereditary myotonia (HM) is a genetic disorder that occurs due to mutations in the chloride channel and results in delayed relaxation of the skeletal muscles. HM has been described in 12 dog breeds, and in five of them, molecular studies of this disorder were performed and mutations in the CLCN1 gene were described. In this study, an affected American Bulldog with HM clinically characterized by muscle hypertrophy, myotonic discharges, and nondystrophic myotonia with a warm-up phenomenon was evaluated, and the candidate canine CLCN1 gene was sequenced. The molecular analysis revealed a frameshift mutation NM_001003124.2:c.436_437insCTCT that resulted in a frameshift and a premature stop codon NP_001003124.1:pTyr146SerfsTer49 . Two aberrant alternative CLCN1 transcripts were observed in an affected dog, the expected transcript with the 4 bp insertion, NM_001003124.2:r.436_437insctct, and an unexpected transcript containing parts of intron 6 in addition to the insertion in exon 4, NM_001003124.2:[r.436_437insctct;r.774_775ins79]. In conclusion, the frameshift mutation in the CLCN1 gene is associated with autosomal recessive HM in American Bulldog and this study constitutes the first description of the disease in this breed. (c) 2020 Elsevier B.V. All rights reserved. Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Rua Prof Dr Walter Mauricio Correa S-N, BR-18618681 Botucatu, SP, Brazil Univ Fed Goias, Sch Vet & Anim Sci, Goiania, Go, Brazil Rodovia Goiania,Km 8 S-N Campus Samambaia, BR-74001970 Goiania, Go, Brazil Univ Ctr UNILEAO, Juazeiro Do Norte, Ceara, Brazil Av Maria Leticia Leite Pereira S-N, BR-63040405 Juazeiro Do Norte, CE, Brazil CALE Anim Surg & Specialized Diagnost Lab, Jundiai, SP, Brazil Rua Italia,106 Jardim Bonfiglioli, BR-13207280 Jundiai, SP, Brazil Rua Parana 330, BR-75709240 Catalao, Go, Brazil Univ Fed Uberlandia, Sch Vet Med, Uberlandia, MG, Brazil Av Mato Grosso 3289,Bloco 2S, BR-38405314 Uberlandia, MG, Brazil Univ Estadual Paulista, Inst Biotechnol, Botucatu, SP, Brazil Alameda Tecomarias S-N, BR-18607440 Botucatu, SP, Brazil Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Univ Estadual Paulista, Inst Biotechnol, Botucatu, SP, Brazil

Made available in DSpace on 2020-12-12T02:39:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-01 Hereditary nephropathy is a primary progressive glomerular disease in dogs associated with the c.115A>T mutation in the COL4A4 gene in English cocker spaniel (ECS) dogs. The disease is inherited in an autosomal recessive manner. Hereditary nephropathy has been described in this breed since the late 1940s. To date, there are no data on the prevalence of this disease in Brazil, so the aim of this study was to evaluate the allelic frequency of this mutation in ECS dogs in this country. The DNA samples were purified from blood samples or buccal swabs from 221 ECS dogs. Fragments of the DNA containing the mutation were amplified by PCR and submitted to direct gene sequencing. The allele frequency of the mutation was 0.9%. The presence of the mutation in the ECS dog population in Brazil reveals the importance of performing the genotyping tests in these dogs as a method of diagnosing the disease and identifying heterozygous animals, aiming to reduce clinical cases of disease through mating. São Paulo State University (Unesp), School of Veterinary Medicine and Animal Science, Rua Prof. Dr. Walter Mauricio Correa, s/n São Paulo State University (Unesp), School of Veterinary Medicine and Animal Science, Rua Prof. Dr. Walter Mauricio Correa, s/n

Mutations in the CLCN1 gene are the primary cause of non-dystrophic Hereditary Myotonia in several animal species. However, there are no reports of Hereditary Myotonia in pigs to date. Therefore, the objective of the present study was to characterize the clinical and molecular findings of Hereditary Myotonia in an inbred pedigree. The clinical, electromyographic, histopathological, and molecular findings were evaluated. Clinically affected pigs presented non-dystrophic recessive Hereditary Myotonia. Nucleotide sequence analysis of the entire coding region of the CLCN1 gene revealed the absence of the exons 15 and 16 in myotonic animals. Analysis of the genomic region flanking the deletion unveiled a large intragenic deletion of 4,165 nucleotides. Interestingly, non-related, non-myotonic pigs expressed transcriptional levels of an alternate transcript (i.e., X2) that was identical to the deleted X1 transcript of myotonic pigs. All myotonic pigs and their progenitors were homozygous recessive and heterozygous, respectively, for the 4,165-nucleotide deletion. This is the first study reporting Hereditary Myotonia in pigs and characterizing its clinical and molecular findings. Moreover, to the best of our knowledge, Hereditary Myotonia has never been associated with a genomic deletion in the CLCN1 gene in any other species.

Made available in DSpace on 2020-12-12T01:52:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Four causative mutations (D1, D2, D3*, and D4) of chondrodysplastic dwarfism have been described in the equine aggrecan (ACAN) gene. Homozygotes for one of these mutations and heterozygotes for any combination of these mutations exhibit the disproportionate dwarfism phenotype. However, no case description of homozygotes for D4 (D4/D4) has been reported in the literature, to our knowledge. We report 2 Miniature horses with the genotype D4/D4 in the ACAN gene. Clinically, the 2 dwarfs had a domed head that was large compared to the rest of the body, mandibular prognathism, and short and bowed limbs, mainly in the proximal region of the metatarsal bones. Radiographic examination revealed contour irregularities of the subchondral bone in the long bones and confirmed mandibular prognathism; histopathology revealed irregular chondrocyte organization. To determine the genotypes of the horses, we performed DNA extraction from white blood cells, PCR, and Sanger sequencing. Genotyping demonstrated that these 2 animals had the D4/D4 genotype in the ACAN gene. The D4/D4 dwarfs were clinically similar to animals with the other ACAN genotypes reported for this disease. Identification of heterozygous animals makes mating selection possible and is the most important control measure to minimize economic losses and casualties. São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science FAPESP: 2016/24025-0 FAPESP: 2016/24767-7 FAPESP: 2018/11365-3

Made available in DSpace on 2019-10-06T16:08:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 This article describes the clinical and electromyographic findings of neuromyotonia in a 19-month-old male crossbred Quarter Horse that presented with stiffness and muscle asymmetry in the hind limbs as well as sacrococcygeal, paravertebral, and gluteal myokymia. An electromyographic study showed spontaneous continuous muscle fiber activity with high-frequency discharges, fibrillations, positive sharp waves, fasciculation potentials, and complex repetitive discharges. Histological examination of the gluteal muscle showed a mixed neurogenic and myopathic pattern. The findings are consistent with neuromyotonia. Department of Veterinary Clinical Sciences School of Veterinary Medicine and Animal Sciences Sao Paulo State University (Unesp) School of Veterinary Medicine and Animal Sciences Federal University of Mato Grosso do Sul School of Veterinary Medicine Federal University of Minas Gerais Department of Neurology Psychology and Psychiatry School of Medicine of Botucatu Sao Paulo State University (Unesp) Federal University of Pelotas Department of Veterinary Clinical Sciences School of Veterinary Medicine and Animal Sciences Sao Paulo State University (Unesp) Department of Neurology Psychology and Psychiatry School of Medicine of Botucatu Sao Paulo State University (Unesp)

Made available in DSpace on 2018-12-11T17:18:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Administration of progesterone (P4) after estradiol is usually performed to prepare non-cyclic mares as embryo recipients. However, there are successful pregnancy reports after embryo transfer in non-cyclic mares treated only with progestins. The objective of this study was to evaluate endometrial gene expression and immunostaining for estrogen receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in seasonal anestrous mares treated with long acting P4 (LA P4). Endometrial tissue from eight seasonal anestrous mares was collected immediately before administration of 1.5 g of LA P4 and five days after. The receptors protein expression was evaluated by immunohistochemistry and the percentage of the immunostained area was determined by ImageJ software. Transcripts abundance for ERα ERβ and PR were determined by RT-qPCR. Blood samples were collected daily to measure plasma P4 concentrations. Protein expression for ERα was greater (p < 0.05) after LA P4 administration, although gene expression was not affected by treatment (p > 0.05). There was no difference for ERβ protein expression (p = 0.07) and ERβ gene expression was reduced (p < 0.05) after treatment. Gene and protein expression for PR was not altered (p > 0.05). In conclusion, endometrial PR and ERα expression patterns after LA P4 administration were similar to those previously found in protocols using estradiol prior to LA P4 to prepare non-cyclic mares as embryo recipients. UNESP-Univ. Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNITRI- Centro Universitário do Triângulo UNESP-Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP- Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science UNESP-Univ. Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNESP-Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP- Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science FAPESP: 2012/00298-7

Made available in DSpace on 2018-12-11T17:17:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) On a farm in southern Brazil, four lambs in a flock of 300 Texel sheep were born with bilateral blindness. They revealed bilateral occlusion of the eyelids and were unresponsive to external visual stimuli and were disoriented when walking. A post-mortem examination revealed bilateral occlusion of the eyelids and, microscopically, total or partial absence of the lens was observed. Blood samples from 20% of the herd (61/300) were sent for molecular analysis. The four affected lambs were homozygous (C/C) for a mutation (single-nucleotide polymorphism c.338G > C) in the PITX3 gene, and 26.2% (16/61) of the herd, including the two ewe and two ram parents, were heterozygous (G/C). Based on clinical, pathological and molecular analysis, it was possible to determine the existence of hereditary microphthalmia in Texel lambs associated with the c.338G > C SNP in the PITX3 gene for the first time in Brazil. The result of this study provides a warning to veterinarians and breeders, emphasizing the importance of considering this disease in the differential diagnosis of congenital diseases in the Brazilian Texel flock. Faculdade de Medicina Veterinária Universidade Federal do Rio Grande do Sul (UFRGS) Departamento de Patologia Veterinária, Av. Bento Gonçalves 9090 Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista (UNESP) Campus Botucatu Departamento de Clínica Veterinária, Rua Prof. Doutor Walter Mauricio Correa s/n Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista (UNESP) Campus Botucatu Departamento de Clínica Veterinária, Rua Prof. Doutor Walter Mauricio Correa s/n CAPES: 1328442

Made available in DSpace on 2019-10-06T15:35:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-04-01 Palicourea marcgravii (Rubiaceae) is considered the most important toxic plant affecting livestock farming in Brazil. This study describes an outbreak of spontaneous poisoning by P. marcgravii in sheep. Forty-nine sheep died while showing clinical signs of sudden death after having access to the plant. During the necropsy, P. marcgravii leaves were observed in the rumens of the animals. Histopathological analysis demonstrated hydropic degeneration of the kidneys. Monofluoracetate concentration obtained in Palicourea samples was 0.53% (5.3 μg/mg). These findings collectively indicate that the affected animals died from poisoning due to P. marcgravii. São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Poisonous Plant Research Laboratory Agricultural Research Service United States Department of Agriculture, 1150 E. 1400 N. Escola de Veterinaria e Zootecnia Universidade Federal de Goias (UFG) São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science

Made available in DSpace on 2018-12-11T17:11:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-07-15 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) To test the hypothesis that the administration of 2.5 mg of estradiol benzoate (EB) followed by 1500 mg of long acting progesterone (LA P4) causes similar uterine changes and molecular dynamics in anovulatory mares to those observed in cyclic ones, we evaluated the changes of estrogen (ERα and ERβ) and progesterone receptors (PR) in anestrous, transitional and cyclic mares by RT-qPCR and immunohistochemistry. In addition, we evaluated uterine edema, tonus and estrogens and progesterone plasma profile. Endometrial biopsies were taken from anestrous and transitional mares immediately before EB injection, 48 h after EB administration and five days after LA P4 was given. In cyclic mares, biopsies were collected at estrus and at five days after ovulation. Similar estrogen peaks were achieved after the injection of the single EB dose between treated and cyclic groups, as well as maximum uterine edema. Uterine tone was increased to diestrus levels after administration of 1500 mg of LA P4. Changes in relative abundance of transcripts for PR, ERα and ERβ when progesterone stimulated endometrium was compared to estrogen stimulated endometrium were similar between cyclic and non-cyclic treated mares. However, apparent decreased PR in the endometrial glandular epithelium was not observed in non-cyclic mares five days after LA P4 administration as observed at five days after ovulation in cyclic mares. The protocol produced similar endometrial edema, uterine tonus and changes in relative abundance of PR, ERα and ERβ transcripts to those observed in cyclic mares during late estrus and early diestrus, as well as similar estradiol and estrogen conjugate plasma concentrations. UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNESP – Univ. Estadual Paulista Campus de Botucatu UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP – Univ. Estadual Paulista Campus de Botucatu UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNESP – Univ. Estadual Paulista Campus de Botucatu UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP – Univ. Estadual Paulista Campus de Botucatu UNESP – Univ. Estadual Paulista Campus de Botucatu Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science FAPESP: 2012/00298-7

Made available in DSpace on 2020-12-12T00:58:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 Mutations in the CLCN1 gene are the primary cause of non-dystrophic Hereditary Myotonia in several animal species. However, there are no reports of Hereditary Myotonia in pigs to date. Therefore, the objective of the present study was to characterize the clinical and molecular findings of Hereditary Myotonia in an inbred pedigree. The clinical, electromyographic, histopathological, and molecular findings were evaluated. Clinically affected pigs presented non-dystrophic recessive Hereditary Myotonia. Nucleotide sequence analysis of the entire coding region of the CLCN1 gene revealed the absence of the exons 15 and 16 in myotonic animals. Analysis of the genomic region flanking the deletion unveiled a large intragenic deletion of 4,165 nucleotides. Interestingly, non-related, non-myotonic pigs expressed transcriptional levels of an alternate transcript (i.e., X2) that was identical to the deleted X1 transcript of myotonic pigs. All myotonic pigs and their progenitors were homozygous recessive and heterozygous, respectively, for the 4,165-nucleotide deletion. This is the first study reporting Hereditary Myotonia in pigs and characterizing its clinical and molecular findings. Moreover, to the best of our knowledge, Hereditary Myotonia has never been associated with a genomic deletion in the CLCN1 gene in any other species. São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Instituto de Medicina Veterinária Universidade Federal do Pará São Paulo State University (UNESP) Medical School Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University São Paulo State University (UNESP) Institute of Bioscience College of Veterinary Medicine University of Minnesota São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Medical School São Paulo State University (UNESP) Institute of Bioscience

Simple Summary Hereditary bleeding disorders occur in different species due to mutations in genes coding specific hemostatic proteins leading to alterations in their synthesis, or to the production of non-functional proteins which leads to impairment of hemostasis. Some of these disorders have been described in horses, i.e., Von Willebrand disease (VWD), hemophilia A, and Glanzmann’s thrombasthenia (GT). GT is an inherited disease characterized by hemorrhage and has been described in different species including horses of varied breeds (Thoroughbred, Standardbred, Oldenburg, Peruvian Paso, and Quarter Horse). There are two different mutations described in horses a single guanine to cytosine substitution (CGG for CCG) and a 10 base pair deletion in the ITGA2B gene. Abstract Glanzmann’s thrombasthenia (GT) is an autosomal recessive inherited disorder characterized by changes in platelet aggregation, leading to hemorrhage and epistaxis. To date, two independent mutations have been described in horses and associated with this disorder, a point mutation (c.122G > C) and a 10-base-pair deletion (g.1456_1466del) in the Integrin subunit alpha2β gene (ITGA2B) of horses of different breeds (Quarter Horse, Thoroughbred, Oldenburg, and Peruvian Paso). ITGA2B codifies the αIIb subunit of the αIIbβ3 integrin, also termed platelet fibrinogen receptor. Horses with GT have been diagnosed in the USA, Canada, Japan, and Australia. However, there are no studies on the prevalence of GT in horses. The aim of this study is to evaluate the prevalence of the mutations responsible for GT in horses in Brazil. A total of 1053 DNA samples of clinically healthy Quarter Horse (n = 679) and Warmblood horses (n = 374) were used. DNA fragments were amplified by PCR and sequenced. The genotype of each animal was analyzed and compared to the nucleotide sequence of the ITGA2B gene found on GenBankTM. There were no carriers in the analyzed samples, that is, all animals tested were wild type. Therefore, under the conditions in which this study was carried out, it can be inferred that GT seems to be extremely rare in the population of Quarter Horses and Warmbloods in Brazil, although it is not possible to affirm that there are no horses carrying mutated alleles in Brazil.

Made available in DSpace on 2019-10-06T16:47:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-10-01 Aiming to investigate the effects of different hormonal protocols on the endometrium morphometry of anestrous mares, 26 animals were assigned to four different treatment groups: (1) EB2.5LAP4: single dose of 2.5 mg of estradiol benzoate (EB); (2) EB5LAP4: 5 mg of EB in 2 consecutive days; (3) EB10LAP4: 10 mg of EB in three consecutive days, considering that all EB-treated groups received a single dose of 1,500 mg of long-acting progesterone (LA P4) after the single/last EB dose; and (4) LAP4: only 1,500 mg of LA P4. Results were also compared with those found in cyclic mares (control group). Endometrial biopsies were collected before and after the hormonal treatments in anestrous mares, and during estrus and at 5 days after ovulation in cyclic mares (D5). Samples were prepared for histological and histomorphometric analysis. Tissue sections were examined to determine luminal epithelium height (LEH), glandular epithelium height (GEH), endometrial thickness (ET), and glandular density (GD). Similar morphometric changes were observed after EB and P4 were administered to groups EB5LAP4 and EB10LAP4. Five days after LA P4 administration (D5), all the assessed variables were similar between all EB-treated groups. In addition, all variables of the EB-treated groups were similar to the control group on D5. Although most of the LAP4 group variables on D5 were similar to the EB-treated groups (except GD), reduced GD and GEH were found when compared with the control group, demonstrating the importance of estradiol priming before P4 on glandular activity and density. UNIPAC- Faculdade Presidente Antônio Carlos UNESP-Universidade Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP-Universidade Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNESP- Universidade Estadual Paulista Campus de Botucatu Department of Anatomy Biosciences Institute UNESP- Universidade Estadual Paulista Campus de Botucatu Department of Biostatistics Biosciences Institute UNESP-Universidade Estadual Paulista Campus de Botucatu Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP-Universidade Estadual Paulista Campus de Botucatu Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science UNESP- Universidade Estadual Paulista Campus de Botucatu Department of Anatomy Biosciences Institute UNESP- Universidade Estadual Paulista Campus de Botucatu Department of Biostatistics Biosciences Institute

Made available in DSpace on 2020-12-12T01:45:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-10-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Progressive retinal atrophy (PRA) due to the c.5G>A mutation in the progressive rod-cone degeneration (PRCD) gene is an important genetic disease in English cocker spaniel (ECS) dogs. Because the prevalence of this disease has not been verified in Brazil, this study aimed to evaluate the allele frequency of the c.5G>A mutation in the PRCD gene. Purified DNA from 220 ECS dogs was used for genotyping, of which 131 were registered from 18 different kennels and 89 were unregistered. A clinical eye examination was performed in 28 of the genotyped animals; 10 were homozygous mutants. DNA fragments containing the mutation region were amplified by PCR and subjected to direct genomic sequencing. The prcd-PRA allele frequency was 25.5%. Among the registered dogs, the allele frequency was 14.9%; among the dogs with no history of registration, the allele frequency was 41%. Visual impairment was observed in 80% (8/10) of the homozygous mutant animals that underwent clinical eye examination. The high mutation frequency found in this study emphasizes the importance of genotyping ECSs as an early diagnostic test, especially as part of an informed breeding program, to avoid clinical cases of PRA. Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (Unesp) São Paulo State University (Unesp) Medical School Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science São Paulo State University (Unesp) São Paulo State University (Unesp) Medical School CNPq: 100232/2018-3

Made available in DSpace on 2019-10-06T16:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-01. Added 1 bitstream(s) on 2019-10-09T18:35:22Z : No. of bitstreams: 1 S0100-736X2019000300175.pdf: 1184744 bytes, checksum: 2868d347ec2bc61ec70388027949a9be (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Albinism is a genetic disease characterized by deficient melanin production making affected animals more susceptible to skin problems, negatively influencing production systems of the same. In buffalo, a nonsense mutation (c.1431G>A) in the tyrosinase gene was already described, which is responsible for the oculocutaneous albinism buffalo phenotype. However, prevalence studies have never been performed for this anomaly in Brazil. Therefore, the objective of this study was to investigate this mutation in buffalo herd in Brazil. Of the 315 buffalo tested with no albinism phenotype evident, 11 (3.5%) were heterozygous for the mutation and none were mutated homozygous, showing the existence of the albinism gene in buffalo production herds and proving the importance of prevalence studies for hereditary diseases in order to prevent the dissemination of these same genes and their negative productivity consequences. Departamento de Clínica Veterinária Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista (Unesp), Rua Prof. Doutor Walter Mauricio Correas/n, Cx. Postal 560 Instituto de Medicina Veterinária Faculdade de Medicina Veterinária Universidade Federal do Pará (UFPA) Campus de Castanhal, Rodovia BR-316 Km 61 Laboratório Regional de Diagnóstico Faculdade de Veterinária Universidade Federal de Pelotas, Rua Gomes Carneiro 1 Embrapa Clima Temperado Campus Universitário s/n Departamento de Clínica Veterinária Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista (Unesp), Rua Prof. Doutor Walter Mauricio Correas/n, Cx. Postal 560 FAPESP: 2015/23569-4 CAPES: 432/2014

Herpesvirus bovino 5 é um alfaherpesvírus causador de meningoencefalite não supurativa em bovinos. Esta doença possui ocorrência natural em surtos ou casos isolados, associadas a baixa morbidade e alta letalidade. Embora estudos anteriores tenham elucidado aspectos relacionados a patogenia da doença, há uma lacuna de conhecimento relacionado aos eventos moleculares que contribuem para a infecção e replicação do BoHV-5. O objetivo do presente estudo foi determinar a expressão gênica in vitro de genes virais (i.e., alfa, beta e gama) e das células hospedeiras (GAPDH e 18S) durante a infecção considerando diferentes momentos de infecção e quantidade de vírus utilizado. Três genes do BoHV-5 (bICP0, UL9, US4), um gene estrutural (GAPDH) e um gene constitutivo (18S) da célula bovina tiveram suas expressões avaliadas por PCR quantitativa (qPCR). Enquanto os genes virais tiveram sua expressão aumentada ao longo do tempo de infecção, o gene hospedeiro teve sua expressão diminuída, demonstrando a ação do vírus na expressão gênica de células bovinas in vitro. O gene constitutivo 18S teve sua expressão mantida durante todos os momentos do experimento. Nossos resultados claramente demonstraram que o GAPDH não deve ser usado como gene de referência em estudos com infecção por BoHV-5 pois é influenciado pela infecção viral. Entretanto, o 18S rRNA foi constitutivamente expresso e pode ser recomendado para normalização em células bovinas infectadas pelo vírus. A expressão de mRNAviral não foi alterada pela quantidade de vírus usada. Todos os genes virais demonstraram o mesmo padrão de expressão ao longo do tempo de infecção. Nossos resultados trazem importantes diferenças comparando aos estudos clássicos que avaliaram a expressão de genes alfa, beta e gama. Mais estudos são necessários para aumentar o conhecimento da biologia molecular do BoHV-5. Estudo utilizando sequenciamento de última geração (i.e., RNA-seq), usando modelos in vitro e in vivo, aparentam ser o próximo passo lógico para acessar as alterações do transcriptoma do hospedeiro e viral ao longo do curso da infecção. Bovine herpesvirus 5 is an alphaherpesvirus that causes nonsuppurative meningoencephalitis in cattle. This disease occurs naturally in either outbreaks or isolated cases, and exhibits low morbidity and high lethality. Although previous studies elucidated crucial aspects involved in the pathogenesis of the disease, there is a paucity of information regarding the molecular events contributing to infection and replication of BoHV-5. The objective of the present study was to determine the in vitro gene expression pattern of BoHV-5 (e.g., alpha, beta, and gamma genes) and host cells genes (GAPDH and 18S) over time utilizing different quantities of inoculated virus. Three BoHV-5 genes (bICP0, UL9, US4) and one structural bovine cell gene had their expression accessed by real-time PCR. While the expression of BoHV-5 genes increased during the course of infection, GAPDH gene expression decreased in the host cells, evidencing the effect of viral infection on the expression of bovine cell genes. The 18S ribosomal RNA (rRNA) gene was constitutively expressed throughout BoHV-5 infection. Our data clearly demonstrates that GAPDH gene should not be used as a reference gene in studies of BoHV-5 infection because it was influenced by viral infection. However, 18S rRNA was constitutively expressed and, therefore, is recommended for normalization of BoHV-5 infection studies in bovine cells. The expression of viral genes transcripts was not altered by increasing number of viral particles added to the culture. All viral genes included here demonstrated the same expression pattern over time and there was no difference in the expression of viral genes among the various time points. Our data show important differences comparing to classical studies regarding herpesvirus alpha, beta, and gamma genes expression. More research is necessary to improve our understanding about the BoHV-5 biology during infection. Studies employing next-generation sequencing (i.e., RNA-seq), using both in vitro and in vivo models, would be the next logical step to grasp the virus and host cell’s transcriptome changes over the course of infection.

Made available in DSpace on 2018-12-11T17:31:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Diarrhea is a major clinical problem affecting foals up to 3 months of age. The aim of this study was to identify enteric microorganisms involved in monoinfections and coinfections and the associated virulence factors in healthy and diarrheic foals. Diarrheic (D) (n=56) and nondiarrheic (ND) foals (n=60) up to three months of age were studied. Fecal samples were analyzed for identification of infectious agents (microbiological culturing, molecular techniques, and microscopic analyses). Escherichia coli fimH (30% versus 25%), Salmonella spp. (25% versus 7%), Strongyloides westeri (25% versus 25%), Clostridium perfringens type A (21% versus 10%), E. coli ag43 (20% versus 35%), Strongylus (11% versus 18%), and vapA-positive Rhodococcus equi (5% versus 2%) were the most frequent enteric pathogens detected in D and ND foals, respectively. The frequency of toxin A-positive C. perfringens was significantly increased in the D (p=0.033) compared with the ND animals. R. equi strains harboring virulent plasmids were also identified (VapA 85-kb type I and VapA 87-kb type I) in D and ND foals. Coinfections were observed in 46% of the D and 33% of the ND foals. Our results demonstrate the great diversity of enteric pathogens, virulence factors, and coinfections involved in enteric infections of foals. Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP) Department of Preventive Veterinary Medicine Federal University of Minas Gerais (UFMG) Department of Genetics Evolution and Bioagents Campinas State University (Unicamp) Department of Preventive Veterinary Medicine University of S�o Paulo Department of Animal Hygiene Kitasato University Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP)

Made available in DSpace on 2020-12-12T02:22:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) In this retrospective study, clinical records of nine horses with a diagnosis of Bothrops envenomation were investigated. The accidents were classified as severe (5/9), moderate (2/9), or mild (2/9) according to the adapted bothropic snakebite severity score (BSSS). All snakebites were on the head region. The main clinical signs were local edema, blood coagulation disorders, and respiratory distress. The whole-blood clotting time (WBCT) was prolonged in all horses, and five horses presented with uncoagulable blood. All horses received specific snake antivenom according to the BSSS (six vials for severe, four vials for moderate, and two vials for mild accidents), and emergency tracheotomy was required in six horses because of respiratory distress. One horse died after eight days of hospitalization, whereas the others were discharged after nine days of hospitalization. The BSSS plus the WBCT were useful in determining the prognosis and the amount and frequency of antivenom therapy. Snakebite accidents are emergency cases; therefore, rapid and efficient therapeutic intervention will reflect positively on the prognosis. São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science São Paulo State University (Unesp) Center for the Study of Venoms and Venomous Animals (CEVAP) São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinical Science São Paulo State University (Unesp) Center for the Study of Venoms and Venomous Animals (CEVAP) FAPESP: 18/25765-3